Data for the co-expression and purification of human recombinant CaMKK2 in complex with calmodulin in Escherichia coli
DATA IN BRIEF
Authors: Gerner, Lisa; Munack, Steffi; Temmerman, Koen; Lawrence-Doerner, Ann-Marie; Besir, Hueseyin; Wilmanns, Matthias; Jensen, Jan Kristian; Thiede, Bernd; Mills, Ian G.; Morth, Jens Preben
Abstract
Calcium/calmodulin-dependent kinase kinase 2 (CaMKK2) has been implicated in a range of conditions and pathologies from prostate to hepatic cancer. Here, we describe the expression in Escherichia coli and the purification protocol for the following constructs: full-length CaMKK2 in complex with CaM, CaMKK2 'apo', CaMKK2 (165-501) in complex with CaM, and the CaMKK2 F267G mutant. The protocols described have been optimized for maximum yield and purity with minimal purification steps required and the proteins subsequently used to develop a fluorescence -based assay for drug binding to the kinase, "Using the fluorescent properties of STO-609 as a tool to assist structure function analyses of recombinant CaMKK2" [1]. Crown Copyright (C) 2016 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/40).
CAMKK2, Regulated by Promoter Methylation, is a Prognostic Marker in Diffuse Gliomas
CNS NEUROSCIENCE & THERAPEUTICS
Authors: Liu, Da-Ming; Wang, Hong-Jun; Han, Bo; Meng, Xiang-Qi; Chen, Ming-Hui; Yang, Dong-Bo; Sun, Ying; Li, Yong-Li; Jiang, Chuan-Lu
Abstract
Aims: To explore the expression, methylation pattern, the prognostic value, and the biological consequences of CAMKK2 in gliomas. Methods: The expression and methylation pattern of CAMKK2 was inferred and validated from mRNA expression profile (N = 866) and methylation profile (N = 426) of glioma tissue samples, and independent samples were used for further validation by IHC and pyrosequencing. To explore the function of CAMKK2 in gliomas, in vitro studies, colony formation assays and migration and invasion assays were performed. Results: We found the upregulation of CAMKK2 in high-grade glioma samples was associated with promoter hypomethylation. An elevated expression of CAMKK2 was associated with worse prognosis. By in vitro assays, we demonstrated that CAMKK2 could promote cell migration, invasion, and proliferation. Conclusions: The expression level of CAMKK2 could be regulated by promoter methylation. CAMKK2 serves as a prognostic marker in gliomas and could be a potential therapeutic target in gliomas.